A simple method for the evaluation of receptor binding capacity of modified cytokines.
We have developed a flow cytometric method to evaluate the binding of interleukin-2 analogues to receptors. The method relies on competition for binding between a fluorescein-conjugated monoclonal antibody (MoAb) directed against the human interleukin-2 receptor alpha chain (fluorescein isothiocyana...
| Main Authors: | , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
1993
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| _version_ | 1797104226106605568 |
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| author | Chianelli, M Signore, A Hicks, R Testi, R Negri, M Beverley, P |
| author_facet | Chianelli, M Signore, A Hicks, R Testi, R Negri, M Beverley, P |
| author_sort | Chianelli, M |
| collection | OXFORD |
| description | We have developed a flow cytometric method to evaluate the binding of interleukin-2 analogues to receptors. The method relies on competition for binding between a fluorescein-conjugated monoclonal antibody (MoAb) directed against the human interleukin-2 receptor alpha chain (fluorescein isothiocyanate (FITC) anti-IL-2R) and the test protein. IL-2R positive cells are incubated with FITC-anti-IL-2R MoAb in the presence of native IL-2 or IL-2 iodinated by either the chloramine-T or the lactoperoxidase-glucose-oxidase method. The binding of IL-2 is indicated by decreased fluorescence. This method is suitable for measuring the binding capacity of modified IL-2 molecules and avoids the need for radioactive tracers. It provides a simple and reproducible technique, which can be extended readily to the study of the receptor binding capacity of cytokines conjugated with toxins, drugs or other molecules. |
| first_indexed | 2024-03-07T06:30:53Z |
| format | Journal article |
| id | oxford-uuid:f5f59685-d2ac-4a9c-8cee-8f3ab1f2d590 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T06:30:53Z |
| publishDate | 1993 |
| record_format | dspace |
| spelling | oxford-uuid:f5f59685-d2ac-4a9c-8cee-8f3ab1f2d5902022-03-27T12:31:20ZA simple method for the evaluation of receptor binding capacity of modified cytokines.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:f5f59685-d2ac-4a9c-8cee-8f3ab1f2d590EnglishSymplectic Elements at Oxford1993Chianelli, MSignore, AHicks, RTesti, RNegri, MBeverley, PWe have developed a flow cytometric method to evaluate the binding of interleukin-2 analogues to receptors. The method relies on competition for binding between a fluorescein-conjugated monoclonal antibody (MoAb) directed against the human interleukin-2 receptor alpha chain (fluorescein isothiocyanate (FITC) anti-IL-2R) and the test protein. IL-2R positive cells are incubated with FITC-anti-IL-2R MoAb in the presence of native IL-2 or IL-2 iodinated by either the chloramine-T or the lactoperoxidase-glucose-oxidase method. The binding of IL-2 is indicated by decreased fluorescence. This method is suitable for measuring the binding capacity of modified IL-2 molecules and avoids the need for radioactive tracers. It provides a simple and reproducible technique, which can be extended readily to the study of the receptor binding capacity of cytokines conjugated with toxins, drugs or other molecules. |
| spellingShingle | Chianelli, M Signore, A Hicks, R Testi, R Negri, M Beverley, P A simple method for the evaluation of receptor binding capacity of modified cytokines. |
| title | A simple method for the evaluation of receptor binding capacity of modified cytokines. |
| title_full | A simple method for the evaluation of receptor binding capacity of modified cytokines. |
| title_fullStr | A simple method for the evaluation of receptor binding capacity of modified cytokines. |
| title_full_unstemmed | A simple method for the evaluation of receptor binding capacity of modified cytokines. |
| title_short | A simple method for the evaluation of receptor binding capacity of modified cytokines. |
| title_sort | simple method for the evaluation of receptor binding capacity of modified cytokines |
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